Sugar esters are widely used in the food, cosmetic, detergent and phar-maceutical industry. The preparation and it’s dynamics of glucose stearic acid esters (GSAE) by lipase-catalyzed reaction were studied in this work. The available lipase of catalytic synthesis GSAE in non-aqueous media was screened from 7 commercial lipases. To facilitate these reactions, a certain volume of glucose, stearic acid, lipase, molecular sieve 3A, solvents etc were added in 50 ml tapered bottles, respectively. The results showed that lipase Novozyme 435 proved to be efficient catalysts of enzymatic preparation GSAE, acetone was appropriate solvent, and Novozyme 435 could keep high activity in acetone. And excessive water in the reaction system obviously decreased the activity of Novozyme 435 and the yield rate of GSAE, thus it was very necessary to add defined amount moisture absorber to remove the water of substrates, solvent and reaction process. The optimized esterification reaction conditions with 0.08 g Novozyme 435, 0.8 g 3A molecular sieve, the ratio of stearic acid and glucose = 3:1, 10 ml acetone, 150r/min shaking, 45℃ reaction temperature, 5 h reaction time were good for making the produce GSAE in non-aqueous media. In the optimized condition, the maximum reaction rate Vm was 0.749 μmol/min?g, Km was 0.0203 mmol/L and the GSAE yield rate was up to 64.11%.